Effect of CuS reinforcement on the mechanical, water vapor barrier, UV-light barrier, and antibacterial properties of alginate-based composite films.

Alginate-based functional nanocomposite films were prepared by the incorporation of copper sulfide nanoparticles (CuSNP) using a solution casting method and characterized their properties. The optical, mechanical, water vapor barrier, surface wetting, and thermal stability properties of the nanocomposite films were tested. The antibacterial activity of the alginate/CuSNP composite film was evaluated against foodborne pathogenic bacteria, E. coli and L. monocytogenes. The morphological test showed that the CuSNP were evenly distributed in the alginate matrix. The addition of 0.5 wt% of CuSNP improved the UV-barrier, hydrophobicity, mechanical strength, and water vapor barrier properties of the alginate-based film; however, the thermal stability was not significantly influenced. The alginate/CuSNP nanocomposite film showed evident antibacterial activity against the Gram-negative (E. coli) bacteria, but only slightly slowed the growth rate of the Garm-positive (L. monocytogenes) bacteria. The alginate/CuSNP nanocomposite films with significantly improved physical and functional properties have a high potential for active food packaging applications.

High performance of alginate/polyvinyl alcohol composite film based on natural original melanin nanoparticles used as food thermal insulating and UV-vis block.

Light is a major factor in promoting food aging and deterioration, especially for ultraviolet (UV) light. Herein, bioinspired dopamine-melanin solid nanoparticles with strong absorption at a wide range of 200-2500 nm were first incorporated into alginate/polyvinyl alcohol to fabricate film materials in this work for UV-vis block, and this also brings excellent thermal insulating properties to the materials. In addition, in order to obtain a material with excellent performance, particles of uniform size of about 100 nm are obtained by fractional centrifugation. It was found the mechanical, UV-vis block and thermal insulating properties were improved significantly compared with the control samples. This study provides a strategy to design a non-polluting, biodegradable, biocompatible film with excellent mechanical properties that can be used in UV-vis barriers and has potential applications in thermal insulating materials for food preservation.

Dual Crosslinked Methacrylated Alginate Hydrogel Micron Fibers and Tissue Constructs for Cell Biology.

As an important natural polysaccharide biomaterial from marine organisms, alginate and its derivatives have shown great potential in the fabrication of biomedical materials such as tissue engineering, cell biology, drug delivery, and pharmaceuticals due to their excellent biological activity and controllable physicochemical properties. Ionic crosslinking is the most common method used in the preparation of alginate-based biomaterials, but ionic crosslinked alginate hydrogels are prone to decompose in physiological solution, which hinders their applications in biomedical fields. In this study, dual crosslinked alginate hydrogel microfibers were prepared for the first time. The ionic crosslinked methacrylated alginate (Alg-MA) hydrogel microfibers fabricated by Microfluidic Fabrication (MFF) system were exposed to ultraviolet (UV) light and covalent crosslink between methacrylate groups avoided the fracture of dual crosslinked macromolecular chains in organizational environment. The chemical structures, swelling ratio, mechanical performance, and stability were investigated. Cell-encapsulated dual crosslinked Alg-MA hydrogel microfibers were fabricated to explore the application in tissue engineering for the first time. The hydrogel microfibers provided an excellent 3D distribution and growth conditions for cells. Cell-encapsulated Alg-MA microfibers scaffolds with functional 3D tissue structures were developed which possessed great potential in the production of next-generation scaffolds for tissue engineering and regenerative medicine.

Near-infrared light-responsive alginate hydrogels based on diselenide-containing cross-linkage for on demand degradation and drug release.

A biodegradable, near-infrared (NIR) - responsive hydrogel is one of the most promising strategies as a remotely triggered drug carrier. In this study, novel NIR-responsive hydrogels based on alginate structures were prepared for controllable drug release. The hydrogels were formed rapidly by reacting norbornene-functionalized alginates and tetrazine cross-linkers containing diselenide bonds via inverse electron demand Diels-Alder click chemistry. In order to manipulate their properties, we prepared hydrogels with various cross-linking densities. NIR sensitive indocyanine green (ICG) and a drug, doxorubicin (DOX) were incorporated in the hydrogel matrix during gelation. The hydrogels showed a suppressed release profile under physiological conditions, while NIR light triggered a rapid release of DOX. Under NIR-light irradiation, ICG generated reactive oxygen species which could decompose diselenide bonds in the hydrogel matrix, inducing the gel-sol transition and release of entrapped DOX. The degradation of hydrogels could be also controlled by the ratio of the precursors.

Ultrasonic Generation of Pulsatile and Sequential Therapeutic Delivery Profiles from Calcium-Crosslinked Alginate Hydrogels.

Control over of biological processes can potentially be therapeutically regulated through localized biomolecular deliveries. While implantable hydrogels can provide localized therapeutic deliveries, they do not traditionally provide the temporally complex therapeutic delivery profiles required to regulate complex biological processes. Ionically crosslinked alginate hydrogels have been shown to release encapsulated payloads in response to a remotely applied ultrasonic stimulus, thus potentially enabling more temporally complex therapeutic delivery profiles. However, thorough characterizations of how different types of therapeutic payloads are retained and ultrasonically released need to be performed. Additionally, the impact of potentially disruptive ultrasonic stimulations on hydrogel structure and temperature need to be characterized to better understand what range of ultrasonic signals can be used to trigger release. To perform these characterizations, calcium-crosslinked alginate hydrogels were loaded with various model macromolecules (dextrans), chemotherapeutics, and protein signaling factors and exposed to a variety of single-pulse and multi-pulse ultrasonic signals at various amplitudes and durations. In response to single-pulsed ultrasonic exposures, quantifications of molecular release, degree of gel erosion, and increase in hydrogel temperature revealed that the ultrasonic stimulations required for statistically significant therapeutic deliveries often eroded and heated the gels to unacceptable levels. However, multi-pulse ultrasonic exposures were shown to achieve significant amounts of therapeutic release while keeping gel erosion and temperature increase at modest levels. Finally, experiments were performed demonstrating that ultrasonic stimulation could be used to generate drug release profiles shown to have potential therapeutic benefits (e.g., pulsatile and sequential anticancer delivery profiles). This work underscores the potential of using ultrasonically responsive polymeric hydrogels for providing on-demand control over more complex therapeutic deliver profiles and enhancing drug delivery strategies in cancer therapies and beyond.

Influence of chitosan-sodium alginate pretreated with ultrasound on the enzyme activity, viscosity and structure of papain.

BACKGROUND: Ultrasound treatment has been shown to be an effective technique for improving the activity of immobilized enzymes. However, its mechanism is unclear. RESULTS: The effect of ultrasonic pretreated chitosan-sodium alginate (CHI-ALG) on the enzymatic activity of papain was investigated via a single factor (temperature, time, frequency, power) experiment. The maximum relative enzyme activity of papain was observed when it was mixed with ultrasound pretreated CHI-ALG at 135 kHz, 0.25 W cm-2 and 50 °C for 20 min, during which the relative activity increased by 72.14% compared to untreated CHI-ALG. Viscosity analysis of papain mixed with CHI-ALG pretreated and untreated with ultrasound revealed that stronger association interactions between the polymers were formed compared to the untreated sample. Fluorescence and circular dichroism spectra indicated that the ultrasonic pretreatment of CHI-ALG increased the number of tryptophan on the papain surface and also increased the content of α-helix by 6.97% and decreased the content of ß-sheet by 3.45% compared to the untreated solution. CONCLUSION: The results of the present study indicate that papain combined with CHI-ALG pretreated with the appropriate ultrasound could be effective technique for improving the activity of immobilized enzymes as a result of changes in its structure and intermolecular interactions. It is important to extend the application of CHI-ALG gel in the immobilized enzyme industry. © 2016 Society of Chemical Industry.

Synthesis and characterization of photo-crosslinkable 4-styryl-pyridine modified alginate.

In this article photo-crosslinkablestyryl-pyridine modified alginate (ASP-Alg) was prepared and entirely investigated utilizing different instrumental techniques such as Elemental analysis, Fourier transform infrared (FTIR),(13)C and (1)H nuclear magnetic resonance (NMR), ultraviolet-visible light (UV-vis), X-ray diffraction (XRD) spectra and scanning electron microscope (SEM). Upon irradiation in the UV region, the casted ASP-Alg membranes were cross-linked through the [2π+2π] cycloaddition reaction of the inserted photo-active styryl pyridine moieties. Both cross-linking density and kinetics were monitored by examining the UV-vis light spectra of the irradiated membrane at predetermined time intervals and the obtained results were found to fit with the second order mathematical kinetic model, revealing the performance of the cross-linking via bimolecular [2π+2π] cycloaddition reaction. Also, the swelling behaviors along with biodegradability were also studied, and the results indicated the decrease of the swelling ratio and degradation rate by increasing the cross-linking density. Moreover, the mechanical properties were also examined under both wet and dry conditions.

Light-responsive iron(III)-polysaccharide coordination hydrogels for controlled delivery.

Visible-light responsive gels were prepared from two plant-origin polyuronic acids (PUAs), alginate and pectate, coordinated to Fe(III) ions. Comparative quantitative studies of the photochemistry of these systems revealed unexpected differences in the photoreactivity of the materials, depending on the polysaccharide and its composition. The roles that different functional groups play on the photochemistry of these biomolecules were also examined. Mannuronic-rich alginates were more photoreactive than guluronic acid-rich alginate and than pectate. The microstructure of alginates with different mannuronate-to-guluronate ratios changed with polysaccharide composition. This influenced the gel morphology and the photoreactivity. Coordination hydrogel beads were prepared from both Fe-alginate and Fe-pectate. The beads were stable carriers of molecules as diverse as the dye Congo Red, the vitamin folic acid, and the antibiotic chloramphenicol. The photoreactivity of the hydrogel beads mirrored the photoreactivity of the polysaccharides in solution, where beads prepared with alginate released their cargo faster than beads prepared with pectate. These results indicate important structure-function relationships in these systems and create guidelines for the design of biocompatible polysaccharide-based materials where photoreactivity and controlled release can be tuned on the basis of the type of polysaccharide used and the metal coordination environment.

Influence of amino acids, buffers, and ph on the γ-irradiation-induced degradation of alginates.

Alginate-based biomaterials and medical devices are commonly subjected to γ-irradiation as a means of sterilization, either in the dry state or the gel (hydrated) state. In this process the alginate chains degrade randomly in a dose-dependent manner, altering alginates' material properties. The addition of free radical scavenging amino acids such as histidine and phenylalanine protects the alginate significantly against degradation, as shown by monitoring changes in the molecular weight distributions using SEC-MALLS and determining the pseudo first order rate constants of degradation. Tris buffer (0.5 M), but not acetate, citrate, or phosphate buffers had a similar effect on the degradation rate. Changes in pH itself had only marginal effects on the rate of alginate degradation and on the protective effect of amino acids. Contrary to previous reports, the chemical composition (M/G profile) of the alginates, including homopolymeric mannuronan, was unaltered following irradiation up to 10 kGy.

Effects of gamma irradiation and moist heat for sterilization on sodium alginate.

Polysaccharides, such as alginates, are already being used as carriers for drug delivery. The physicochemical and biological properties of alginates may be affected via irradiation and thermal treatments. To explore and compare effects of two kinds of sterilization methods, gamma irradiation and moist heat, on sodium alginate (SA), physicochemical and biological properties of SA powder and solutions were investigated after sterilization. Human umbilical vein endothelial cells (HUVEC) was used to assess the cytotoxicity of the SA after sterilization. The research showed that 25 kGy gamma ray can effectively sterilize microorganism. Both gamma irradiation and moist heat hardly affect the native pH of SA. Compared to irradiation sterilization, moist heat sterilization showed smaller changes in intrinsic viscosity for all SA samples and lead to less glycosidic bond breaking of SA powders. The moist heat sterilization can cause the main chain scission and double bonds formation of the SA solutions. Cytotoxicity studies demonstrated that sterilized SA powers and SA solutions treated by gamma ray sterilization can increase the viability of HUVEC. However, SA solutions treated by moist heat sterilization were found to present severe cytotoxicity. The research results may provide interesting future advancements toward the development of SA-based products for biomedical applications.

Radiolytically depolymerized sodium alginate improves physiological activities, yield attributes and composition of essential oil of Eucalyptus citriodora Hook.

Eucalyptus citriodora Hook. is highly valued for its citronellal-rich essential oil (EO) extracted from its leaves. Hence, escalated EO production of eucalyptus is the need of hour. Marine polysaccharides (sodium alginate) are processed through gamma radiation of particular intensity, to obtain the irradiated sodium alginate (ISA). A pot experiment was conducted to study the effect of foliar application of ISA on growth, biochemical, physiological, EO yield and composition of E. citriodora. The treatments were applied as: foliar spray of deionized water only (control), seed soaked with ISA (90 mg L(-1)) and foliar spray of ISA with 30, 60, 120 and 240 mg L(-1). The treatment 6 (spray of ISA at 120 mg L(-1)) showed the highest value for most of the parameters studied. It also enhanced the EO content (33.3%), EO yield (86.7%), citronellal content (63.4%) and citronellal yield (205.5%) as compared to the control.

Photo-activated ionic gelation of alginate hydrogel: real-time rheological monitoring of the two-step crosslinking mechanism.

We examine the gelation of alginate undergoing ionic crosslinking upon ultraviolet (UV) irradiation using in situ dynamic rheology. Hydrogels are formed by combining alginate with calcium carbonate (CaCO3) particles and a photoacid generator (PAG). The PAG is photolyzed upon UV irradiation, resulting in the release of free calcium ions for ionic crosslinking. The viscous and elastic moduli during gelation are monitored as a function of the UV irradiation intensity, exposure time, alginate concentration, and the ratio between alginate and calcium carbonate. Gel time decreases as irradiation intensity increases because a larger concentration of PAG is photolyzed. Interestingly, dark curing, the continuing growth of microstructure in the absence of UV light, is observed. In some instances, the sample transitions from a solution to a gel during the dark curing phase. Additionally, when exposed to constant UV irradiation after the dark curing phase, samples reach the same plateau modulus as samples exposed to constant UV without dark curing, implying that dark curing does not affect the gelation mechanism. We believe the presence of dark curing is the result of the acidic environment persisting within the sample, allowing CaCO3 to dissociate, thereby releasing free Ca(2+) ions capable of binding with the available appropriate ionic blocks of the polymer chains. The growth of microstructure is then detected if the activation barrier has been crossed to release sufficient calcium ions. In this regard, we calculate a value of 30 J that represents the activation energy required to initiate gelation.

Terminal sterilization of alginate hydrogels: efficacy and impact on mechanical properties.

Terminal, or postprocessing, sterilization of composite biomaterials is crucial for their use in wound healing and tissue-engineered devices. Recent research has focused on optimizing traditional biomaterial formulations to create better products for commercial and academic use which incorporate hydrophobic compounds or secondary gel networks. To use a hydrogel in a clinical setting, terminal sterilization is necessary to ensure patient safety. Lyophilization, gamma-irradiation, and ethylene oxide treatment all have negative consequences when applied to alginate scaffolds for clinical use. Here, we aim to find alternative terminal sterilization methods for alginate and alginate-based composite hydrogels which maintain the structure of composite alginate networks for use in biomedical applications. A thorough investigation of the effect of common sterilization methods on swollen alginate-based hydrogels has not been reported and therefore, this work examines autoclaving, ethanol washing, and ultraviolet light as sterilization techniques for alginate and alginate/Pluronic® F68 composite hydrogels. Preservation of structural integrity is evaluated using shear rheology and analysis of water retention, and efficacy of sterilization is determined via bacterial persistence within the hydrogel. Results indicate that ethanol sterilization is the best method of those investigated because ethanol washing results in minimal effects on mechanical properties and water retention and eliminates bacterial persistence. Furthermore, this study suggests that ethanol treatment is an efficacious method for terminally sterilizing interpenetrating networks or other composite hydrogel systems.

Preparation of oligoalginate plant growth promoter by γ irradiation of alginate solution containing hydrogen peroxide.

Degraded alginate compounds with molecular weights of 7-26, 40-77, or 11-26 kDa were obtained by γ irradiation, hydrogen peroxide (5% H(2)O(2)) treatment, or a combination treatment involving ionizing radiation and H(2)O(2), respectively. The 14 kDa oligoalginate, prepared by the combined method, promoted the growth of mustard greens and lettuce at an optimal concentration of 75 mg/L. The growth promotion effects of the oligoalginate prepared by γ irradiation in the presence of H(2)O(2) were statistically equivalent to those of the oligoalginate prepared by γ irradiation only. The combination of γ irradiation and H(2)O(2) reduced the required irradiation dosage by a factor of 9 relative to the oligoalginate produced by γ irradiation only. The combination treatment (γ irradiation/H(2)O(2)) may be carried out on a large scale at low cost to produce oligoalginate for use as a plant growth promoter in agricultural industries.

Anisotropic structure of calcium-induced alginate gels by optical and small-angle X-ray scattering measurements.

It was more than 50 years ago that an appearance of birefringence in alginate gels prepared under cation flow was reported for the first time, however, the anisotropic structure of the alginate gel has not been studied in detail. In the present study, anisotropic Ca-alginate gels were prepared within dialysis tubing in a high Ca(2+)-concentration external bath, and optical and small-angle X-ray scattering (SAXS) measurements were performed to characterize the structure of the gel. The observations of the gel with crossed polarizers and with circular polarizers revealed the molecular orientation perpendicular to the direction of Ca(2+) flow. Analyses of the SAXS intensity profiles indicated the formation of rod-like fibrils consisting of a few tens of alginate molecules and that the anisotropy of the gel was caused by the circumferential orientation of the large fibrils. From the observed asymmetric SAXS pattern, it was found that the axis of rotational symmetry of the anisotropic structure was parallel to the direction of Ca(2+) flow. The alignment factor (A(f)) calculated from the SAXS intensity data confirmed that the orientation of the fibrils was perpendicular to the direction of Ca(2+) flow.

Photosensitive controlled release with polyethylene glycol-anthracene modified alginate.

Covalent modification of alginate with polyethylene glycol-conjugated anthracene molecules has the potential to both stabilize the alginate and act as a photosensitive crosslinker. Release studies with Coomassie Blue show lengthy release times from the alginate photogels that extend past 70 days with, for example, 17% versus 27% release at 1750 h (73 days) for photogels with and without 365-nm UV light treatment for 30 min at 10 mW/cm(2) in the initial release period. Photocrosslinking of the photogels after loading effectively "locks" in drug compounds to control their release. Effective crosslinking densities and controls of polyethylene glycol-crosslinked alginate and physically crosslinked calcium alginate gels suggest strong interactions between Coomassie Blue and both alginate and anthracene. Photogels containing anthracene-capped star-polyethylene glycol show increased photosensitivity with modified release profiles. Ultimately, the covalent modification of alginate with photoactive crosslinkers has the potential to produce a long-term, photosensitive, controlled release system.

Preparation and evaluation of hydrogel-composites from methacrylated hyaluronic acid, alginate, and gelatin for tissue engineering.

Hydrogels are three-dimensional water-insoluble hydrophilic natural or synthetic polymer networks made up of crosslinked water-soluble polymers. The purpose of this study was to develop and directly compare photo crosslinked hydrogels on the basis of pure gelatin, alginate and hyaluronic acid as well as their blends. The functionalization of starting materials with methacrylate moieties was evaluated by 1H-NMR spectroscopy. Hydrogels were prepared from methacrylates by photo cross-linking using UV light. The effect of changing the hydrogel composition was quantified through examination of hydrogel swelling behavior and rheological properties. In addition, the viability and adhesion of neonatal rat cardiomyocytes (NRCM) seeded onto the hydrogels was examined by in vivo imaging of NRCM-mediated scaffold contraction as well as by histological evaluation after immunostaining. Biological testing showed good biocompatibility and cell survival in the presence of all materials discussed. Adhesion of cells could only be observed in the presence of gelatin. Blends of gelatin, alginate and hyaluronic acid are promising candidates for the generation of non-toxic, biocompatible hydrogel scaffolds for tissue engineering. Variation of individual compound ratios in the blends can be used for a precise control of mechanical properties and may allow wide-ranging uses in various tissue engineering applications with different mechanical requirements.

Stiffness of photocrosslinked RGD-alginate gels regulates adipose progenitor cell behavior.

Adipose progenitor cells (APCs) are widely investigated for soft tissue reconstruction following tumor resection; however, the long-term success of current approaches is still limited. In order to develop clinically relevant therapies, a better understanding of the role of cell-microenvironment interactions in adipose tissue regeneration is essential. In particular, the effect of extracellular matrix (ECM) mechanics on the regenerative capability of APCs remains to be clarified. We have used artificial ECMs based on photocrosslinkable RGD-alginate to investigate the adipogenic and pro-angiogenic potential of 3T3-L1 preadipocytes as a function of matrix stiffness. These hydrogels allowed us to decouple matrix stiffness from changes in adhesion peptide density or extracellular Ca(2+) concentration and provided a physiologically relevant 3D culture context. Our findings suggest that increased matrix rigidity promotes APC self-renewal and angiogenic capacity, whereas, it inhibits adipose differentiation. Collectively, this study advances our understanding of the role of ECM mechanics in adipose tissue formation and vascularization and will aid in the design of efficacious biomaterial scaffolds for adipose tissue engineering applications.

Hemostasis for severe hemorrhage with photocrosslinkable chitosan hydrogel and calcium alginate.

In patients with severe hemorrhage, complications such as shock or death may occur if the patient is not treated appropriately and expeditiously. To create a hemostat kit for severe hemorrhage, ultraviolet light irradiation was applied to photocrosslinkable chitosan hydrogel and calcium alginate. As a hemorrhage model, the femoral arteries and veins of anesthetized rats were cut. Hemodynamics and hematological parameters including red blood cell (RBC) count, hemoglobin concentration, hematocrit, white blood cell (WBC) count, and platelet count, and serum parameters including aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were measured as a marker of hemostasis. In rats for which no procedure was used, death occurred within 30 min. By using the hydrogel hemostat, the survival rate rose to 75% or more. RBC count, hemoglobin, hematocrit, and platelet levels were not significantly changed for 3 days. WBC count increased 1 day after hemostasis. AST and ALT increased 1 day after hemostasis, but it decreased 3 days later. The photocrosslinkable chitosan hydrogel and calcium alginate were biodegraded at 3 and 28 days, respectively, by neutrophils and keratinocyte chemoattractant.

Patterning alginate hydrogels using light-directed release of caged calcium in a microfluidic device.

This paper describes a simple reversible hydrogel patterning method for 3D cell culture. Alginate gel is formed in select regions of a microfluidic device through light-triggered release of caged calcium. In the pre-gelled alginate solution, calcium is chelated by DM-nitrophen (DM-n) to prevent cross-linking of alginate. After sufficient UV exposure the caged calcium is released from DM-n causing alginate to cross-link. The effect of using different concentrations of calcium and chelating agents as well as the duration of UV exposure is described. Since the cross-linking is based on calcium concentration, the cross-linked alginate can easily be dissolved by EDTA. We also demonstrate application of this capability to patterned microscale 3D co-culture using endothelial cells and osteoblastic cells in a microchannel.